Point of Care Detection of Oral Pathogens

 描述(申请人提供)：有许多致病微生物在口腔内引起疾病或在那里可以检测到。其中包括三种重要病毒：人类免疫缺陷病毒(HIV)、人乳头瘤病毒(HPV)和单纯疱疹病毒(HSV)。艾滋病毒感染和疾病是可以治疗的，但费用高昂，而且仍与相当大的发病率和死亡率有关。人乳头瘤病毒是口咽癌的主要病因，单纯疱疹病毒引起常见的口腔病变。所有这些病原体都可以通过接触唾液传播。这不是艾滋病毒的常见传播途径，但它仍然令人担忧，因为艾滋病毒会导致艾滋病，艾滋病是一种致命的疾病。因此，希望有一种用于检测HIV、HPV和HSV感染的简单、成本效益高的护理点(POC)设备。我们建议使用蛋白质微阵列通过捕获抗原来检测HIV，并基于与微阵列中病毒蛋白的结合来检测针对每种病毒的抗体。抗原发现公司(ADI)在蛋白质微阵列的开发、生产和销售方面拥有丰富的经验。我们现在销售来自各种致病病毒、细菌和原生动物的17个反应蛋白抗原微阵列，其中包括一个HIV-1阵列，其中包含来自五个最常见亚型的蛋白质。此外，ADI已经从HPV、HSV 1型和2型的11种血清型中分离出所有基因，并将它们插入到表达载体中，用于生产蛋白质。为了开发一种有用的POC设备来检测唾液中的HIV以及针对HIV、HPV和HSV的抗体，我们在该项目的第一阶段有以下具体目标：1.创建包含所有常见的HIV-1、HPV和HSV类型的所有蛋白质的蛋白质组芯片。稍后，我们将扩大微阵列以包括其他口腔病原体2.开发一种使用蛋白质组微阵列对唾液中口腔病原体抗体进行敏感和特异筛选的方案。在第二阶段，将开发一种微流控装置，使分析过程自动化。3.建立灵敏、特异的HIV抗原捕获检测方法。在第二阶段，将开发针对HPV、HSV和其他口腔病原体的类似检测方法，并将使用微流控设备实现检测过程的自动化。

 DESCRIPTION (provided by applicant): There are many pathogenic organisms that cause disease in the oral cavity or that can be detected there. Among these are three important viruses, human immunodeficiency virus (HIV), human papilloma virus (HPV) and herpes simplex virus (HSV). HIV infection and disease is treatable but is costly and is still associated with considerable morbidity and mortality. HPV is the primary cause of oropharyngeal cancer and HSV causes common oral lesions. All of these pathogens can be spread by contact with saliva. This is not a common route of transmission for HIV, but it is nevertheless a concern since HIV causes AIDS which is a fatal disease. It would therefore be desirable to have a simple, cost-effective point-of-care (POC) device for the detection of infection by HIV, HPV and HSV. We propose to use a protein microarray to detect HIV by antigen capture and to detect antibodies to each virus based on binding to viral proteins in the microarray. Antigen Discovery Incorporated (ADi) has considerable experience in the development, production and marketing of protein microarrays. We now sell seventeen reactive protein antigen microarrays from a variety of pathogenic viruses, bacteria and protozoa including an HIV-1 array with proteins from the five most common subtypes. Moreover, ADi has already isolated all the genes from eleven serotypes of HPV, HSV types 1 and 2 and inserted them into an expression vector for production of proteins. In order to develop a useful POC device for the detection of HIV as well as antibodies against HIV, HPV and HSV in saliva, we have the following specific aims for the first phase of this project: 1. Create a proteomic microarray containing all proteins from all common types of HIV-1, HPV and HSV. Later we will expand the microarray to include other oral pathogens 2. Develop a protocol for sensitive and specific screening for antibodies to oral pathogens in saliva using a proteomic microarray. In Phase II a microfluidic device will be developed to automate the assay procedure. 3. Develop a sensitive and specific microarray-based antigen-capture detection assay for HIV. In Phase II similar assays will be developed for HPV, HSV and other oral pathogens and the assay procedure will be automated using a microfluidic device.